LOCUS       sequence-75909-e         707 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Golden Gate entry vector at position 2, LacZ, final entry vector for
            the assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 707)
  AUTHORS   Kirchmaier S, Lust K, Wittbrodt J
  TITLE     Golden GATEway cloning--a combinatorial approach to generate fusion 
            and recombination constructs.
  JOURNAL   PLoS One. 2013 Oct 7;8(10):e76117. doi: 
            10.1371/journal.pone.0076117.
  PUBMED    24116091
REFERENCE   2  (bases 1 to 707)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
            date: "2013-10-7"; volume: "8(10):e76117. doi"; pages: " 
            10.1371/journal.pone.0076117"
COMMENT     Sequence Label: pGGEV_2'_XcmI-LacZ
FEATURES             Location/Qualifiers
     source          1..707
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        27..45
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     CDS             complement(83..466)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSYLGDTIEYSSYASNALGALPYGRPAGGREFTSDIEFPR
                     PPWRPGACDVGPNSPYSESYYNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEA
                     RTDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     232..254
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     246..263
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     247..263
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     270..289
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        270..288
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      319..328
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      323..332
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(426..444)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(462..478)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(462..478)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(475..497)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    486..502
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(510..516,517..534,535..540))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    555..576
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 tgcctgggga cgtcggagca agcttgattt aggtgacact atagaggtct caggctccgg
       61 atccagcggt ggcgtggcgc gcttaatgcg ccgctacagg gcgcgtccat tcgccattca
      121 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
      181 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
      241 gacgttgtaa aacgacggcc agtgaattgt aatacgactc actatagggc gaattgggcc
      301 cgacgtcgca tgctcccggc cgccatggcg gccgcgggaa ttcgatatca ctagtgaatt
      361 cgcggccgcc tgcaggtcga ccatatggga gagctcccaa cgcgttggat gcatagcttg
      421 agtattctat agtgtcacct aaatagcttg gcgtaatcat ggtcatagct gtttcctgtg
      481 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
      541 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
      601 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
      661 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactc
//
